Экспериментально-клиническое обоснование использования комплекса диквертина и аскорбиновой кислоты при монополярной коагуляции яичников

There has been conducted a study of morphofunctional ovary changes at monopolar coagulation using histological, histochemical and ultra-microscopic techniques, and there have been evaluated hemorheologic factors and state of lipid peroxidation system (LPS) in postsurgical period. It has been proved that electrocoagulation stimulates the follicle growth for a short time but damages the generative and endocrine ovary apparatus and causes evident hemodynamic changes in perifocal zone. At the same time the raise of blood viscosity and erythrocyte aggregation and LPS activation are occurring. Possessing hemorheologic and antioxidant properties ascovertin produces a protective effect on generative ovary apparatus and microcirculation in it. 48 women after laparoscopic operations on ovaries with the use of electrocoagulation have been examined prospectively. The women of the basic group took the diquertin and ascorbic acid complex prior to and after the surgery in addition to the basic therapy. There has been made an evaluation of LPS factors prior to and after the surgery, computed kymopertubation results and hormonic state in a 1 month after the surgery, frequency of pregnancy beginning. Prescription of diquertin and ascorbic acid complex under the developed procedure is pathogenetically reasonable, it allows to increase the quality of treatment after laparoscopic operations on ovaries and encourages the preservation of reproductive function of a female organism.Проведено исследование морфофункциональных изменений яичников при монополярной коагуляции с использованием гистологических, гистохимических и ультрамикроскопических методик, оценены гемореологические показатели и состояние системы перекисного окисления липидов (ПОЛ) в послеоперационном периоде. Показано, что электрокоагуляция кратковременно стимулирует рост фолликулов, но оказывает повреждающее действие на генеративный и эндокринный аппарат яичников, вызывает выраженные гемодинамические изменения в перифокальной области. Одновременно происходит повышение вязкости крови, агрегации эритроцитов и активация системы ПОЛ. Обладая гемореологическими и антиоксидантными свойствами, асковертин оказывает протективное влияние на генеративный аппарат яичников и состояние микроциркуляции в органе. Проведено проспективное исследование 48 женщин, которым были выполнены лапароскопические операции на яичниках с использованием электрокоагуляции. Женщины основной группы до и после операции на фоне базисной терапии получали комплекс диквертина и аскорбиновой кислоты. Оценивали показатели системы ПОЛ до и после операции, данные компьютерной кимопертубации и гормональный статус через 1 мес после операции, частоту наступления беременности. Назначение комплекса диквертина и аскорбиновой кислоты по разработанной методике является патогенетически обоснованным, позволяет повысить качество лечения после лапароскопических операций на яичниках, способствует сохранению репродуктивной функции женского организма

Участие активных форм кислорода в регуляции Са2+ - активируемых К + -каналов эритроцитов

In this study, we investigated the effects of preincubation with the reactive oxygen species-generating system xanthine oxidase/xanthine on Ca2+-dependent potassium permeability of erythrocyte membrane. The increase of intracellular calcium concentration in presence of calcium ionophore A23187 led to erythrocyte membrane hyperpolarization due to opening of Ca2+-activated potassium channels. Erythrocyte membrane potential was recorded via measurement of pH of the incubation medium in presence of prothonophore. Incubation of erythrocytes with xanthine (100 µmol)/ xanthine oxidase (10 mU/ml) mixture resulted in significant loss of amplitude and rate of hyperpolarization response and also loss the rate of membrane potential restoration. These effects can be caused by hydrogen peroxide, one of products of reaction of xanthine oxidase/xanthine.Изучен эффект предынкубации эритроцитов с активными формами кислорода, генерируемыми системой ксантиноксидаза - ксантин, на Са2+-зависимую калиевую проницаемость мембраны этих клеток. Увеличение внутриклеточной концентрации кальция в присутствии кальциевого ионофора А23187 вело к гиперполяризации эритроцитарной мембраны вследствие открывания Са2+-зависимых калиевых каналов. Мембранный потенциал эритроцитов был зарегистрирован благодаря измерению рН среды инкубации в присутствии протонофора. Инкубация эритроцитов в присутствии ксантина (100 мкмоль) и ксантиноксидазы (10 мU/мл) приводило к значительному снижению амплитуды и скорости развития гиперполяризации, а также к уменьшению скорости восстановления мембранного потенциала. Эти эффекты могут быть вызваны перекисью водорода - одним из продуктов ксантиноксидазной реакции

Поражение печени при хроническом вирусном гепатите С на фоне опийной наркомании: морфологические аспекты

To ascertain the severity of opium-induced liver injury for chronic viral hepatitis C (HCV) patients with detection of morphological characters of hepatocyte impact by opiates.Needle biopsy of liver and the light microscopy of biopsy material were made both for isolated HCV and HCV+opiomania patients.Liver injury (hepatitis activity, fibrosis stage) for HCV patients against the background of opiomania does not differ from the case of isolated HCV. Use of alcohol by opium addicts even in very small doses (10—15 g per day) cause the development of significantly more severe liver injury. Fibrosis of central vein was found significantly more often for HCV against the background of opiomania.Fibrosis of central vein is a differential-diagnostic criterion of being opiates as an etiological factor of liver injury. Opiates as an isolated toxic factor does not cause severe liver injury for HCV patients unlike the use of alcohol even in small doses.Проведено исследование тяжести поражения печени при хроническом вирусном гепатите С (ХВГ С) на фоне опийной наркомании с выявлением морфологических феноменов воздействия опиатов на гепатоциты.Пациентам с ХВГ С и с течением ХВГ С на фоне опийной наркомании проведена пункционная биопсия печени и световая микроскопия биоптатов.При ХВГ С на фоне опийной наркомании тяжесть поражения печени (активность гепатита, стадия фиброза) не отличается от изолированного течения ХВГ С. Употребление опиатными наркоманами даже крайне малых, так называемых негепатотоксичных, доз алкоголя (10—15 г этанола в сутки) обусловливает развитие достоверно более тяжелого поражения печени. При системном употреблении опиатов, по данным морфологического исследования биоптатов, достоверно чаще встречается фиброз центральных вен.Установлено, что фиброз центральных вен является характерным морфологическим признаком воздействия опиатов на печень. Опиаты как изолированный токсический фактор не приводят к более тяжелому поражению печени при ХВГ С, что происходит при употреблении даже малых доз алкоголя

Характеристика эпидемиологических и молекулярных взаимоотношений аллергических и гельминтных болезней в эндемическом очаге описторхоза

To elucidate the molecular mechanisms of O. felineus impact into phenotypic variability of allergic diseases in the opisthorchis endemic region, we studied 104 patients with opisthorchosis, 92 patients with atopic bronchial asthma, 52 patients with a combination of both diseases, and 120 healthy persons. Standard clinical, immunological, and genetic methods were used. An association of opisthorchis invasion with the improvement of lung function signs and bronchial hyperreactivity was found. It was established, that IL-4-dependent mechanisms of atopy were suppressed by O. felineus antigens, in particular due to hyperproduction of IL-10 and transforming growth factor-beta. However, IL-5-dependant mechanisms were supported. A phenomenology of the cytokine gene differential expression was established, disclosing the molecular basis of the immune system function in diseases with polar immune response in the helminth endemic region.С целью исследования молекулярных механизмов вклада O. felineus в фенотипическую изменчивость аллергических болезней в регионе, эндемичном по описторхозу, обследовано 104 больных описторхозом, 92 - атопической бронхиальной астмой, 52 больных с сочетанием бронхиальной астмы и описторхоза и 120 здоровых лиц. Использованы стандартные клинические, иммунологические и генетические методы. Показана ассоциация описторхозной инвазии с повышенной частотой аллергических болезней и высоким уровнем общего иммуноглобулина E. Установлена связь описторхоза с улучшением показателей функции легких и бронхиальной гиперреактивности. Выявлено, что IL-4-зависимые механизмы супрессируются антигеном O. felineus, в частности, за счет гиперпродукции IL-10 и трансформирующего ростового фактора-β (TGFβ), а IL-5-зависимые механизмы иммунного ответа - поддерживаются. Установлена феноменология дифференциальной экспрессии генов цитокинов, раскрывающая молекулярные основы функционирования иммунной системы при болезнях с полярным иммунным ответом в эндемическом регионе

L1pred: A Sequence-Based Prediction Tool for Catalytic Residues in Enzymes with the L1-logreg Classifier

To understand enzyme functions, identifying the catalytic residues is a usual first step. Moreover, knowledge about catalytic residues is also useful for protein engineering and drug-design. However, to experimentally identify catalytic residues remains challenging for reasons of time and cost. Therefore, computational methods have been explored to predict catalytic residues. Here, we developed a new algorithm, L1pred, for catalytic residue prediction, by using the L1-logreg classifier to integrate eight sequence-based scoring functions. We tested L1pred and compared it against several existing sequence-based methods on carefully designed datasets Data604 and Data63. With ten-fold cross-validation, L1pred showed the area under precision-recall curve (AUPR) and the area under ROC curve (AUC) of 0.2198 and 0.9494 on the training dataset, Data604, respectively. In addition, on the independent test dataset, Data63, it showed the AUPR and AUC values of 0.2636 and 0.9375, respectively. Compared with other sequence-based methods, L1pred showed the best performance on both datasets. We also analyzed the importance of each attribute in the algorithm, and found that all the scores contributed more or less equally to the L1pred performance

ИНСУЛИН И ИНСУЛИНОРЕЗИСТЕНТНОСТЬ: НОВЫЕ МОЛЕКУЛЫ-МАРКЕРЫ И МОЛЕКУЛЫ-МИШЕНИ ДЛЯ ДИАГНОСТИКИ И ТЕРАПИИ ЗАБОЛЕВАНИЙ ЦЕНТРАЛЬНОЙ НЕРВНОЙ СИСТЕМЫ

The review summarizes current data on the role of insulin in the regulation of t glucose metabolism in the central nervous system at physiologic and pathologic conditions. For many years, the brain has been considered as an insulin-independent organ which utilizes glucose without insulin activity. However, it is become clear now that insulin not only regulates glucose transport and metabolism, but also has modulatory efftects in impact on excitability, proliferation and differentiation of brain progenitor cells, synaptic plasticity and memory formation, secretion of neurotransmitters, apoptosis. We have critically reviewed literature information and our own data on the role of insulin and insulin resistance in neuron-glia metabolic coupling, regulation of NAD+ metabolism and action of NAdependent enzymes, neurogenesis, brain development in (patho)physiological conditions. The paper clarifies interrelations between alterations in glucose homeostasis, development of insulin resistance and development of neurodegeneration (Alzheimer's disease and Parkinson's disease), autism, stroke, and depression. We discuss the application of novel molecular markers of insulin resistance (adipokines, α-hydroxybutyrate, BDNF, insulin-regulated aminopeptidase, provasopressin) and molecular targets for diagnostics and treatment of brain disorders associated with insulin resistance.В статье рассматриваются вопросы, связанные с ролью инсулина в обмене глюкозы в центральной нервной системе в физиологических условиях и при развитии нейродегенеративных заболеваний и инсулинорезистентности. Долгие годы головной мозг считался инсулиннезависимым органом, способным утилизировать глюкозу без участия инсулина. В настоящее время инсулину принадлежит не только роль регулятора транспорта глюкозы и ее метаболизма, но и модулятора таких процессов, как электровозбудимость нейронов, пролиферация и дифференцировка прогениторных клеток, синаптическая пластичность, формирование памяти, секреция нейротрансмиттеров, апоптоз. В настоящем обзоре критически проанализирована современная литература, с учетом собственных данных, о роли инсулина и инсулинорезистентности в регуляции нейрон-глиального метаболического сопряжения, поддержании гомеостаза НАД+ и регуляции активности НАД+ -зависимых ферментов, нейрогенеза и развития мозга в (пато)физиологических условиях. Раскрываются причинно-следственные связи между нарушением гомеостаза глюкозы, развитием инсулинорезистентности и нейродегенеративными заболеваниями (болезнь Альцгеймера и Паркинсона), аутизмом, острым нарушением мозгового кровообращения, депрессией. Обсуждается применение новых молекул-маркеров инсулинорезистентности (адипокины, α-гидроксибутират, BDNF, инсулинрегулируемая аминопептидаза, провазопрессин) и молекул-мишеней для диагностики и терапии заболеваний головного мозга, ассоциированных с развитием инсулинорезистентности.

Improved siRNA/shRNA Functionality by Mismatched Duplex

siRNA (small interfering RNA) and shRNA (small hairpin RNA) are powerful and commonly used tools in biomedical research. Currently, siRNAs are generally designed as two 21 nt strands of RNA that include a 19 nt completely complementary part and a 2 nt overhang. However, since the si/shRNAs use the endogenous miRNA machinery for gene silencing and the miRNAs are generally 22 nt in length and contain multiple internal mismatches, we tested if the functionality can be increased by designing the si/shRNAs to mimic a miRNA structure. We systematically investigated the effect of single or multiple mismatches introduced in the passenger strand at different positions on siRNA functionality. Mismatches at certain positions could significantly increase the functionality of siRNAs and also, in some cases decreased the unwanted passenger strand functionality. The same strategy could also be used to design shRNAs. Finally, we showed that both si and miRNA structured oligos (siRNA with or without mismatches in the passenger strand) can repress targets in all individual Ago containing cells, suggesting that the Ago proteins do not differentiate between si/miRNA-based structure for silencing activity

Why Are Outcomes Different for Registry Patients Enrolled Prospectively and Retrospectively? Insights from the Global Anticoagulant Registry in the FIELD-Atrial Fibrillation (GARFIELD-AF).

Background: Retrospective and prospective observational studies are designed to reflect real-world evidence on clinical practice, but can yield conflicting results. The GARFIELD-AF Registry includes both methods of enrolment and allows analysis of differences in patient characteristics and outcomes that may result. Methods and Results: Patients with atrial fibrillation (AF) and ≥1 risk factor for stroke at diagnosis of AF were recruited either retrospectively (n = 5069) or prospectively (n = 5501) from 19 countries and then followed prospectively. The retrospectively enrolled cohort comprised patients with established AF (for a least 6, and up to 24 months before enrolment), who were identified retrospectively (and baseline and partial follow-up data were collected from the emedical records) and then followed prospectively between 0-18 months (such that the total time of follow-up was 24 months; data collection Dec-2009 and Oct-2010). In the prospectively enrolled cohort, patients with newly diagnosed AF (≤6 weeks after diagnosis) were recruited between Mar-2010 and Oct-2011 and were followed for 24 months after enrolment. Differences between the cohorts were observed in clinical characteristics, including type of AF, stroke prevention strategies, and event rates. More patients in the retrospectively identified cohort received vitamin K antagonists (62.1% vs. 53.2%) and fewer received non-vitamin K oral anticoagulants (1.8% vs . 4.2%). All-cause mortality rates per 100 person-years during the prospective follow-up (starting the first study visit up to 1 year) were significantly lower in the retrospective than prospectively identified cohort (3.04 [95% CI 2.51 to 3.67] vs . 4.05 [95% CI 3.53 to 4.63]; p = 0.016). Conclusions: Interpretations of data from registries that aim to evaluate the characteristics and outcomes of patients with AF must take account of differences in registry design and the impact of recall bias and survivorship bias that is incurred with retrospective enrolment. Clinical Trial Registration: - URL: http://www.clinicaltrials.gov . Unique identifier for GARFIELD-AF (NCT01090362)

Improved risk stratification of patients with atrial fibrillation: an integrated GARFIELD-AF tool for the prediction of mortality, stroke and bleed in patients with and without anticoagulation.

OBJECTIVES: To provide an accurate, web-based tool for stratifying patients with atrial fibrillation to facilitate decisions on the potential benefits/risks of anticoagulation, based on mortality, stroke and bleeding risks. DESIGN: The new tool was developed, using stepwise regression, for all and then applied to lower risk patients. C-statistics were compared with CHA2DS2-VASc using 30-fold cross-validation to control for overfitting. External validation was undertaken in an independent dataset, Outcome Registry for Better Informed Treatment of Atrial Fibrillation (ORBIT-AF). PARTICIPANTS: Data from 39 898 patients enrolled in the prospective GARFIELD-AF registry provided the basis for deriving and validating an integrated risk tool to predict stroke risk, mortality and bleeding risk. RESULTS: The discriminatory value of the GARFIELD-AF risk model was superior to CHA2DS2-VASc for patients with or without anticoagulation. C-statistics (95% CI) for all-cause mortality, ischaemic stroke/systemic embolism and haemorrhagic stroke/major bleeding (treated patients) were: 0.77 (0.76 to 0.78), 0.69 (0.67 to 0.71) and 0.66 (0.62 to 0.69), respectively, for the GARFIELD-AF risk models, and 0.66 (0.64-0.67), 0.64 (0.61-0.66) and 0.64 (0.61-0.68), respectively, for CHA2DS2-VASc (or HAS-BLED for bleeding). In very low to low risk patients (CHA2DS2-VASc 0 or 1 (men) and 1 or 2 (women)), the CHA2DS2-VASc and HAS-BLED (for bleeding) scores offered weak discriminatory value for mortality, stroke/systemic embolism and major bleeding. C-statistics for the GARFIELD-AF risk tool were 0.69 (0.64 to 0.75), 0.65 (0.56 to 0.73) and 0.60 (0.47 to 0.73) for each end point, respectively, versus 0.50 (0.45 to 0.55), 0.59 (0.50 to 0.67) and 0.55 (0.53 to 0.56) for CHA2DS2-VASc (or HAS-BLED for bleeding). Upon validation in the ORBIT-AF population, C-statistics showed that the GARFIELD-AF risk tool was effective for predicting 1-year all-cause mortality using the full and simplified model for all-cause mortality: C-statistics 0.75 (0.73 to 0.77) and 0.75 (0.73 to 0.77), respectively, and for predicting for any stroke or systemic embolism over 1 year, C-statistics 0.68 (0.62 to 0.74). CONCLUSIONS: Performance of the GARFIELD-AF risk tool was superior to CHA2DS2-VASc in predicting stroke and mortality and superior to HAS-BLED for bleeding, overall and in lower risk patients. The GARFIELD-AF tool has the potential for incorporation in routine electronic systems, and for the first time, permits simultaneous evaluation of ischaemic stroke, mortality and bleeding risks. CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier for GARFIELD-AF (NCT01090362) and for ORBIT-AF (NCT01165710)